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Abstract: Even though osteoarthritis (OA) is mainly considered as a degradative condition of the articular cartilage, there is increasing body of data demonstrating the involvement of all branches of the immune system. Genetic, metabolic or mechanical factors cause an initial injury to the cartilage resulting in release of several cartilage specific auto-antigens, which trigger the activation of immune response. Immune cells including T cells, B cells and macrophages infiltrate the joint tissues, cytokines and chemokines are released from different kinds of cells present in the joint, complement system is activated, and cartilage degrading factors such as matrix metalloproteinases (MMPs) and prostaglandin E2 (PGE2) are released, resulting in further damage to the articular cartilage. There is considerable success in the treatment of rheumatoid arthritis using anti-cytokine therapies. In OA, however, these therapies did not show much effect, highlighting more complex nature of pathogenesis of OA. This needs the development of more novel approaches to treat OA, which may include therapies that act on multiple targets. Plant natural products have this kind of property and may be considered for future drug development efforts. Here we reviewed the studies implicating different components of the immune system in the pathogenesis of OA. [Copyright &y& Elsevier]

Abstract: α-Crystallin, a predominant protein of the ocular lens, is composed of two subunits, αA and αB. Of these, αB-crystallin has been shown to present widely in non-lenticular tissues while αA-crystallin is largely lens-specific. Although, expression of αB-crystallin is elevated under various stress and pathological conditions, yet its physiological significance remained unknown. Some studies suggest that the expression of αB-crystallin gene is related to oxidative stress. Persistent hyperglycemia during uncontrolled diabetes is known to cause oxidative stress, which has been implicated in various secondary complications of diabetes. Hence, expression of αA- and αB-crystallins in various tissues of streptozotocin (STZ)-induced diabetic Wistar-NIN rats was investigated by RT-PCR and immunoblotting. While expression of αB-crystallin was noted in the wide range of tissues examined in the study, αA-crystallin expression was detected only in lens and retina. Interestingly, αB-crystallin expression was elevated in lens, heart, muscle, and brain, but decreased in adipose tissue of diabetic rats compared to control rats. αA-Crystallin expression was increased in retina of diabetic rat. Increased oxidative stress appears to be a major stimulus for the enhanced expression of αA- and αB-crystallins in the tissues of diabetic rats and elevated expression of α-crystallin may have a protective role against metabolic stress. Interestingly, feeding of curcumin, a dietary antioxidant, to diabetic rats attenuated the enhanced expression of αB-crystallin. The results indicate that elevated expression of α-crystallins in some tissues may have implications in pathophysiology of diabetic complications. [Copyright &y& Elsevier]

Exosomes are secreted from various cells by multivesicular bodies (MVBs) that fuse with the plasma membrane and are involved in the intestinal immune response to maintain intestinal homeostasis. Here, we demonstrate the ultrastructural characteristics of MVBs and their exosomes in immune-related cells of the zebrafish intestine, including goblet cells (GCs), mitochondria-rich cells (MRCs), high endothelial cells (HECs) and lymphocytes. In GCs, MVBs with a low electron density were present under the nucleus. MVBs with exosomes were observed among mucin granules. "Heterogeneous" MVBs were identified within the cytoplasm around mucin granules. MRCs were observed in the intestinal mucosa epithelium, including "open-type" MRCs and "close-type" MRCs. Typical MVBs were identified in these MRCs. MVBs with a variety of exosomes were observed in the HECs of the capillary located in the lamina propria (LP). The HEC basement membrane budded outward to LP cells to form a plurality of basal blebs, later containing a large number of exosomes. MVBs also existed in the LP lymphocytes. A schematic diagram of the ultrastructural distribution of MVBs and their exosomes in the intestinal mucosal immune-related cells was created. Our findings provide cytological evidence for the source and ultrastructural distribution of exosomes within the different intestine cells of zebrafish. Component analysis and immunological functions of exosomes require future study. • "Heterogeneous" MVBs existed in goblet cells of zebrafish intestine. • High endothelial cells secreted exosomes by the way of basal blebs. • MVBs and their exosomes were presented in two-types of mitochondria-rich cells. • Illustrative diagram of MVBs-exosomes in zebrafish intestinal immune-related cells. [ABSTRACT FROM AUTHOR]

Abstract Intercellular communication of gut epithelial cells is critical to gut mucosal homeostasis. Exosomes are important intercellular mediators in communication between cell to cell. Although many literature focus on the immunologic roles in the gut by the exosomes, the biological process of exosomes in the absorptive cells remains unknown. Uncovering the distribution, classification and formation process of multivesicular bodies (MVBs) and their exosomes in the absorptive cells of the zebrafish gut, is urgently needed to establish a platform for immunological research of fish gut exosomes. The expression levels of CD63 and TSG101 were different among the three segments of the gut, and they were enriched at the apex of the mid gut villi. The characteristics of MVBs and their exosomes in the absorptive cells were further revealed by transmission electron microscopy (TEM). Early endosomes (ee) were mainly present in the apical and basal cytoplasm of absorptive cells. Late endosomes (le) were mostly distributed with the supranuclear part of these cells. "Heterogeneous" MVBs were detected underlying the apical membranes of absorptive cells. Many exosomes with some MVB-like structures occurred in the lumen, indicating that the release process was mainly through apical secretion. Various MVBs with exosomes and the endosome-heterogeneous MVB-exosome complex existed widely in the mid gut absorptive cells, concluding that zebrafish as a potential model for in vivo MVBs and their exosomes research. All the results were summarized in a schematic diagram illustrating the morphological characteristics of gut MVBs and their exosomes in zebrafish. Highlights • MVBs and their exosomes were present in the mid gut absorptive cells. • Numerous exosomes with some MVB-like structures released into the gut lumen. • The schematic diagram of in vivo MVBs and their exosomes in the gut of zebrafish. [ABSTRACT FROM AUTHOR]

Abstract It is conceivable that early developing germ cells must across the basal to the luminal region of seminiferous tubules (STs) during spermatogenesis is associated with extensive restructuring of junctional complex. However, very limited information is documented about these junctional complexes in reptiles. In the present study we have determined the localization of inter-Sertoli cell tight junctions (TJ's), protein CLDN11 and gap junction protein Cx43 during spermatogenesis in the testis. In early spermatogenesis, weak immunoreactivity of CLDN11and focal localization of Cx43 was observed around the Sertoli cell in the luminal region, but completely delaminated from the basal compartment of STs. In late spermatogenesis, strong focal to linear localization of CLDN11and Cx43 was detected at the points of contact between two Sertoli cells and around the early stages of primary spermatocytes in the basal compartment of STs. In late spermatogenesis, localization of CLDN11and Cx43 was drastically reduced and seen only around Sertoli cells and spermatogonia near the basal lamina. However, transmission electron microscopy revealed that inter-Sertoli cell tight junctions were present within the basal compartment of STs, leaving the spermatogonia and early primary spermatocytes in the basal region during mid spermatogenesis. Gap junctions were observed between Sertoli cells, and Sertoli cells with spermatogonia and primary spermatocytes throughout spermatogenesis. Moreover, adherens and hemidesmosomes junctions were observed during spermatogenesis. The above findings collectively suggest that the intensity and localization of TJ's and gap junctions vary according to the spermatogenetic stages that might be protected the developing germ cells from own immune response. Highlights • This is the first study provide information about the tight and gap junction's location during spermatogenesis in turtle. • Intensity of tight junctions varies during spermatogenesis according to different stages of developing germ cells. • We determined that the gap junctions are highly associated with the development of inter-Sertoli cell tight junctions. [ABSTRACT FROM AUTHOR]

Despite many studies being conducted over the past few decades, the origin of autophagosomal membranes remains unclear. The present study aimed to uncover the formation process of autophagosomal membranes in hepatocytes of zebrafish ( Danio rerio ), a model organism in medical science. Immunohistochemistry of zebrafish hepatocytes indicated that light chain 3-like protein 2 (LC3-II) is highly active in some hepatocytes, but poorly expressed in others. Under transmission electron microscopy, the amount of autophagosomes (APs) varied in different hepatocytes. When the endoplasmic reticulum (ER) is dispersed in the cytoplasm, few isolation membranes (IMs) and APs were observed. Subsequently, when the ER assembles into a particular “lamellar structure” (LS), IMs arise from it and extend to enwrap the mitochondria. With further aggregation of the ER, the LS developed into an over twenty-layered structure, and mitophagy was more obvious in the hepatocytes and cavities appeared in mitochondria. Finally, most ERs were assembled into several LSs. At this point, mitophagy was most active in the hepatocytes. Thereafter, glycogen and lipid droplet increased gradually, while the LS degenerated and ER scatter increased. Then, the glycogen and lipid droplets dominated the hepatocellular cytoplasm. After suppressing the formation of autophagosomes using 3-Methyladenine (3-MA), the LS could no longer be visualized in the hepatocellular cytoplasm, and mitophagy decreased drastically. Taken together, the results suggested that this LS in the hepatocytes of zebrafish, might be another manifestation of a pre-autophagosomal structure in zebrafish liver, analogous to the omegasome in yeast or the ER-IM complex in mammalian cell lines. Furthermore, selective mitophagy and consequent cyclic utilization of its products were probably relevant to dynamic cycle of the hepatocellular cytoplasm. [ABSTRACT FROM AUTHOR]

To mitigate greenhouse gas (GHG) emissions, different strategies have been proposed, including application of dolomite, crop straw and biochar, thus contributing to cope with the increasing global warming affecting the planet. In the current study, pristine wheat straw biochar (WBC) and magnesium (MgCl 2.6H 2 O) modified wheat straw biochar (MWBC) were used. Treatments included control (CK), two WBC dosages (1% and 2.5%), and two MWBC doses (1% and 2.5%). After 90 days of incubation, WBC and MWBC improved the soil physiochemical properties, being more pronounced with increasing rates of biochar. MWBC 2.5 significantly decreased microbial biomass carbon (MBC), while microbial biomass nitrogen (MBN) increased when both biochar materials (WBC 1 and MWBC 1) were applied at low rate. Compared to control soil, Urease and Alkaline phosphatase activities increased with the increasing rate of WBC and MWBC. The activities of dehydrogenase and β-glucosidase decreased with the WBC and MWBC application, compared to CK. The fluxes of all the three GHGs evaluated (CO 2 , CH 4 and N 2 O) decreased with time for both biochar amendments, while cumulative emission of CO 2 increased by 58% and 45% for WBC, and by 54% and 41% for MWBC, as compared to CK. The N 2 O cumulative emissions decreased by 18 and 34% for WBC, and by 25 and 41% for MWBC, compared to CK, whereas cumulative methane emission showed non-significant differences among all treatments. These findings indicate that Mg-modified wheat straw biochar would be an appropriate management strategy aiding to reduce GHG emissions and improving the physiochemical properties of affected soils, and specifically of the red dry land soil investigated in the current work. • MBC decrease by both biochars while MBN increase at low rate of WBC and MWB. • Urease and phosphatase activity increased while dehydrogenase and β-glucosidase activity decreased. • WBC and MWBC increased cumulative CO2, whereas decreased the N2O emission. [ABSTRACT FROM AUTHOR]

Resistin, an adipocyte secreted cysteine rich hormone has been implicated as molecular link between obesity and type 2 diabetes in a murine model. Although, at the protein level mouse and human resistin show remarkable similarities with respect to conserved cysteine residues, the physiological role of human resistin is not yet clear. In the present study we describe the purification and refolding of human recombinant resistin using two different refolding processes. Gel filtration analysis of protein refolded by both the methods revealed that human recombinant resistin, like mouse resistin, has a tendency to form dimers. Interestingly, dimerization of resistin appears to be mediated by both covalent (disulfide bond mediated) and non-covalent interactions as seen on reducing and non-reducing SDS–PAGE. Circular dichroism spectral analysis revealed that human resistin peptide backbone is a mixture of α-helical and β-sheet conformation with significant amounts of unordered structure, similar to the mouse resistin. It is likely that the first cysteine (Cyst22) of human resistin, which is equivalent to mouse Cyst26, may be involved in stabilizing the dimers through covalent interaction. [Copyright &y& Elsevier]